Luo An, He Jie, Yu Jinpeng, Wu Yong, Harvey Peta J, Kasheverov Igor E, Kudryavtsev Denis S, McIntosh J Michael, Tsetlin Victor I, Craik David J, Zhangsun Dongting, Luo Sulan
Key Laboratory of Tropical Biological Resources of Ministry of Education, Hainan University, Haikou 570228, China; Guangxi Key Laboratory of Special Biomedicine, School of Medicine, Guangxi University, Nanning 530004, China.
Guangxi Key Laboratory of Special Biomedicine, School of Medicine, Guangxi University, Nanning 530004, China.
Int J Biol Macromol. 2024 Jun;271(Pt 1):132472. doi: 10.1016/j.ijbiomac.2024.132472. Epub 2024 May 19.
The two most active disulfide bond isomers of the analgesic αO-conotoxin GeXIVA, namely GeXIVA[1, 2] and GeXIVA[1, 4], were subjected to Asp-scanning mutagenesis to determine the key amino acid residues for activity at the rat α9α10 nicotinic acetylcholine receptor (nAChR). These studies revealed the key role of arginine residues for the activity of GeXIVA isomers towards the α9α10 nAChR. Based on these results, additional analogues with 2-4 mutations were designed and tested. The analogues [T1A,D14A,V28K]GeXIVA[1, 2] and [D14A,I23A,V28K]GeXIVA[1, 4] were developed and showed sub-nanomolar activity for the α9α10 nAChR with IC values of 0.79 and 0.38 nM. The latter analogue had exceptional selectivity for the α9α10 receptor subtype over other nAChR subtypes and can be considered as a drug candidate for further development. Molecular dynamics of receptor-ligand complexes allowed us to make deductions about the possible causes of increases in the affinity of key GeXIVA[1, 4] mutants for the α9α10 nAChR.
对镇痛性αO-芋螺毒素GeXIVA的两种活性最强的二硫键异构体，即GeXIVA[1, 2]和GeXIVA[1, 4]进行天冬氨酸扫描诱变，以确定在大鼠α9α10烟碱型乙酰胆碱受体（nAChR）上发挥活性的关键氨基酸残基。这些研究揭示了精氨酸残基对GeXIVA异构体针对α9α10 nAChR活性的关键作用。基于这些结果，设计并测试了另外具有2至4个突变的类似物。开发出了类似物[T1A,D14A,V28K]GeXIVA[1, 2]和[D14A,I23A,V28K]GeXIVA[1, 4]，它们对α9α10 nAChR表现出亚纳摩尔活性，IC值分别为0.79和0.38 nM。后一种类似物对α9α10受体亚型相对于其他nAChR亚型具有出色的选择性，可被视为进一步开发的候选药物。受体-配体复合物的分子动力学使我们能够推断出关键的GeXIVA[1, 4]突变体对α9α10 nAChR亲和力增加的可能原因。
